Figure S1 Glucocorticoid receptor-green fluorescent protein (GR-GFP) reporter construct.

Transcription

1 Figure S1 Glucocorticoid receptor-green fluorescent protein (GR-GFP) reporter construct. (A) Schematic display of the construct is presented detailing binding sites and regions of interest within the GR promoter including primer and other putative binding sites (GR2-FR, GR-RV), glucocorticoid response element (GRE), activator protein 1 binding site (AP-1), splicing factor 1 binding site (SP-1), estrogen response element (ERE), and transcription start site (TSS). (B) Sequence of putative GR promoter region, complete with specific locations of each putative binding site. Figure S2 Fluorescence images of GR promoter activation in developing embryos. A GR-GFP construct consisting of zebrafish GR promoter region coupled to the GFP coding sequence was injected into embryos (n=9 groups of embryos). No fluorescence was detected at 12 hpf (I), but fluorescence was observed at 15 hpf (II) and 18 hpf (III), while a negative control construct lacking the GR promoter region did not fluoresce (IV). Arrows indicate regions of fluorescence. Figure S3 Putative non-functional glucocorticoid response elements in MyoD and IGF promoter regions. Schematics of putative glucocorticoid response elements (GREs; numbered by starting base pair) upstream of MyoD, IGF1, and IGF2.

2 Figure S1A

3 Figure S1B TCGTAAACATGCTTGCTTTGGAGAGGTTGAGATATATCAATAATGATAAATTGGAGATATTAAATAAAACATAGGG ACAGATATTAGAACACCCTAATGCGTTGGAAGAAATGCAAGCTTAACCAATGACCCCTGTCAAAAACATAGAAAAT CAAT Box AAAGATTAAGTGTAATTGATATTCATTCATTCATTTTCTTTTTGGCTTAGTTCCTTTATTAATCCAGAGTCGCCACAG AATAAACCGCCATTTTATCCAGCGTTTATGCAGCGGATGTCCTTCCAGCTGCAACCCATCTCTGGGAAACATCCATA GRE CATACTCATTCACACTCATACACTACGGACAATTTAGTCTAGCCAATTCACCTGTTTTTGGACTGGAGGAAACCGGA GGAAATCCAAGTAAATGCGGGGAGAACATGCAAAGTCCACACAGAAACGGCAACTGACCCAGCCGAGGCTTGAA AP 1 CCGGCAACCTTCTTGCTGTGAGGCGACAGCACTACTTATTGCGCCACTGTGTCACCCCTGTAATTGACATACACTAT TAATAATCTATAATAGATAAATAAAACTATAGTAATATAACAACTTACTTTATTATTACTATTATGAATGCAGTACTT GACCAGCTAAATTATATTTTGCTTTCATGATCATTTGTCCCTCTGTGTTCTGTTGTTCTGTGTTTTTATGTATCGTGTT TTATATCTATTATTATTGTATTATATTGTTTTTTCTTCTATATTTTGTTGTAGTTTTATAAAATGCAAAAAGCACAAAA AACGTTGAAAAAATTAAATTACAGGTGGAACTCAGAGGATGTTAATCAAACAAACAAATGTACTTCACTGACCTTT CAATTGAATTATTACACTGTAAAATCCAACAGTCAACTTTATCAAATGAAATGAGTGTAGTTAACTCAAAATGTACT GAAAGTTAATTCTACTCATTTGAAAAGAGTTTTGAACTCAGCGGTAAAGGTAATGAGTTAATTAAATACCTCATTAC TTCAACTTAAACGGAGTAAGTTCACAGTACTCATATATATATATATATATAATATAGATTAGTTTAACTCAAATGGTT TGTTGCAATCGGTTTCCTCAAATGGTTTGAGTTGCCTTAACTTACTGGGTTTTACAGTGTATTCCAGCTTGTTAAACC AAAACTAATTAGTCACGTATGCACTATCACTCATTTCACAACAAATGATTAACATACAGATCACCTTTATATCTACAT CGCTCTCTCCATAAAAAACTGTCTCAAATAAATTAAAAGTCATTCTGAATAAAAGAATTTATTTGCCTGTGCCAGTT TTGATTCATCACGGATTATTGTTGCAGTTATAACCGTGTTACTTCTTTATAATGTAGTATGTAGAGTGCAATAAATG CTATCAGATAAAAAAGAAAGACATAATTCAAGGTGACAACAAGAGTCTCATTTAGTGATTCTAATACATAAAGCTC ATTAAAAGTGTTTTTTTTTCTTATCTTAAAACAAAAAGCATCCACTAGGTGGAAGTGTTTAGTTGTCACCTACATGTT TATTTCAGTAATTATCTTCTCTAATGTCATCAATTTAGCTAAACAGCTGAGTCATCACGGAGAAAATGAACACTAGC TGCCATCTTTAAAACCTGAACCTTCAACAAATAATTTCAATTTACCAATGTAATCATTCATTCAACAATTCTAAACGA CGGTTAATCTCTTAACCAACATAGTTATTATAGTTATTAATATTTTTACTTGATTTTTAAAATGAATTATTTCCTTTGC

4 ATTTATTTCATAATAGTTGTCTTTTTTAATATAAATGAAATCTGACATAACAGGCCATTATTTATTTAGTTCTTAACAA TGAATGAAAGTAAGACATAAAATAAAAGTGTTCACGTTTGTTTTCCTATGGTTTAGTCCCTTTTTTCATCAGGAGTC ACCACAGCGGAATGAACTGCCAACTTATCTAGCACATGTTTTACACTGCAGCTGCCCTTCCAGCAGCAACCCATTAC TGAAAACCTCCATACACACTCAAAATTTATCTTTAAAGAACAAACACTCTAAACAACGCTGGTTTGTTTAACTCTAC ATTTGAGTAAAATGTCAACAAGTTGTGTCCACTATGTACATTACTGGTCAAAAGTTTGGGGTCAGATTTTGTACTTT TTTTCTTTTTTTTTTTTTTAAGAAAATGATTCTGTTCATCAAGATGGCATTTATTAAATACAAAAAAAATTTTATTGTT AAATATTTATTAAAAGATTTAATAAATGCTTAGTGGATGTAGTTTGAAATGGAAGTATTACTCCAGTCATTGTTGCT TTTATTACTATTACTATTAATAATATTAAGAACTGTAATAATTAATATTAGATGATTTCTAAAGGATCATATGACTCT GAGAACTGGAGTTATAAAACTGAAAATTCATCTTTAAAATCACTGGAATTAATTATTAAATTAAATTATAAAGTACT TTTGAACAGTTCTTTTATAGTGCAAGAACACTTCACAATTTTAATGTTTGTACTGTATTTTTTATTTAATAAATGTGTC CTTGGTGAGTGAGAAGCATATTTTAAAACATTTAAAAATCCTACTGACCCCAAACTTCTGACCGGAAGTGTATTTCA AAGAATTTTCAGGTGGGTTTGACCATATCTTGTATAAACTGATTGTTGTAACAATGCATAAGCTAAATTCTTTGAAA ERE CAAACATTACATTCTGAGCTCCGGAGGTTTTAAAAGATTTAAATGTTAACATTTCCTTGCGTTTTACACACTCATAAA TGTATAATGGAATAATGTTAAATATTTGATCCCACTTTAAACCCAACATTTTGGTTTGTTCACATTTTACAGAAATGT AGGGTGACGCAAACCATGCGTTGTTTACTTACAAAACCAGCATTAATTCAATAATTCAAATAGTCAAAACAACAAC AACAACAACAACAACAACAACAACAACAGTTTGATAGGCTATAGATAATGCAACAAACAAAAAAGGGAAATTCTA ACAAAGACAGTTTGGTTTTATGCATTCTGTCAAAATAATAAACACAAAGCAACAAAGTAATTGCAATTTGTTTGAA ATATAAACCATAGTATAGCCATTTTTTACTTTTGATTTTCATAATGATCATTGATGCATGAACGAATAATCCTATTTC AAATTACAAATAAAAGTTTTTGAAAAACTTTCTAAATGTATGTACAAGGGAAATGCATAATAAAAGCAGAGAGAG TSS* TATA box AGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGGAGGCTGTGTGATTTGT TTTTATGGTGATATCACCGACTCAGACCCAGATCACACTCTCTCACCCAATGAAGTCACTGTAATAGGAAGCGAGA TGCAATAACTCTGTACTGCCAATAGGTCGCACTGCATGTTGTAAACAGGGTCGCCGCGGACCTTTCTACTGTAATCT CCATTTTGAATGTTTTTATGGTTTAGGGTTTTGT

5 Figure S2

6 Figure S3